Isolating DNA from Gram-Negative Bacteria

descriptionPublicationkeyboard_double_arrow_right Article 01 Jan 2017 English Publisher:Cold Spring Harbor LaboratoryJournal:Cold Spring Harbor Protocols, volume 2,017, page pdb.prot093369 (issn: 1940-3402, eissn: 1559-6095,

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Authors: Michael R, Green; Joseph, Sambrook;

doi: 10.1101/pdb.prot093369

pmid: 28049781

Isolating DNA from Gram-Negative Bacteria

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Abstract

The isolation of DNA from bacteria, described in this protocol, relies upon the use of sodium dodecyl sulfate and proteinase K to lyse the cells. High-molecular-weight DNA is then sheared (to reduce its viscosity and make it more manageable), extracted with phenol:chloroform, and precipitated with isopropanol. DNA isolated according to this procedure ranges from 30 to 80 kb in length.

Keywords

DNA, Bacterial, Phenol, Detergents, Sodium Dodecyl Sulfate, 2-Propanol, Bacteriolysis, Gram-Negative Bacteria, Solvents, Chloroform, Endopeptidase K, Molecular Biology

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